Fuller, 16 Comments 1989 reports that the presence of glycerol in a sample loaded on an electrophoretic gel may cause artifacts, such as distortion of nucleic acid fragments in DNA sequencing gels. The distortion is said to resemble a bulge in the sequencing gel, and renders the 400-600 nucleotide region of the gel unreadable. Fuller states that because of the distortion caused by the presence of glycerol, United States Biochemical Corporation supplies a sequencing enzyme, SEQUENASE Version 2.0 T7 DNA polymerase, at a high enough concentration so that no glycerol distortion occurs when the enzyme is diluted prior to use.
Carninci et al. 18 Nucleic Acids Research 204, 1989 describe a standard sequencing gel system using Tris/Borate/EDTA buffer (TBE). It also describes a discontinuous buffer system using Tris-sulphate and Tris-borate. The Tris-sulphate is used as a running gel buffer, and Tris-borate as a tank buffer.
Richards, et al., 12 Analytical Biochemistry 452, 1965 and Peacock and Dingman, 6 Biochemistry 1818, 1967 describe electrophoresis of ribonucleic acid in polyacrylamide gels and resolution of multiple RNA species by polyacrylamide gel electrophoresis. Richards, et al., describe use of Tris-HCl buffer as well as acetic, cacodylic, diethyl barbituric, and glycyl glycine buffers. Peacock and Dingman describe use of Tris-EDTA and boric acid buffers for electrophoresis. The RNA species are not provided in glycerol-containing samples.
Ansorge and Barker, 9 J. Bioc. Biop. Meth. 33, 1984 describe use of Tris/Tricine buffer and an ammediol system for electrophoresis of Maxam and Gilbert DNA sequencing products. Such products are provided in samples without glycerol.
Brumley and Smith, 19 Nuc. Acid. Res. 4121, 1991 describe use of a borate buffer for a sequencing gel.